A Mixture of Baicalein, Wogonin, and Oroxylin-A Inhibits EMT in the A549 Cell Line via the PI3K/AKT-TWIST1-Glycolysis Pathway.

Non-small cell lung cancer (NSCLC) is a worldwide disease with a high morbidity and mortality rate, which is most derived from its metastasis. Some studies show that the epithelial-mesenchymal transition (EMT) process promotes lung cancer cell migration and invasion, leading to NSCLC metastasis. Total flavonoid aglycones extract (TFAE) isolated from Scutellaria baicalensis was reported to inhibit tumor growth and induce apoptosis. In this study, we found that baicalein, wogonin, and oroxylin-A were the active compounds of TFAE. After reconstructing with these three compounds [baicalein (65.8%), wogonin (21.2%), and oroxylin-A (13.0%)], the reconstructed TFAE (reTFAE) inhibited the EMT process of A549 cells. Then, bioinformatic technology was employed to elucidate the potential pharmacodynamic mechanism network of reTFAE. We identified the relationship between reTFAE and PI3K/Akt signaling pathways, with TWIST1 as the key protein. LY294002, the inhibitor of the PI3K/Akt signaling pathway, and knock-down TWIST1 could significantly enhance the efficacy of reTFAE, with increasing expression of epithelial markers and decreasing expression of mesenchymal markers in A549 cells at the same time. Furthermore, stable isotope dimethyl-labeled proteomics technology was conducted to complement the follow-up mechanism that the EMT-inhibition process may be realized through the glycolysis pathway. In conclusion, we claim that TWIST1-targeted flavonoids could provide a new strategy to inhibit EMT progress for the treatment of NSCLC.

[Impact on the gait time cycle of ischemic stroke in the treatment with yin-yang respiratory reinforcing and reducing needling technique].

OBJECTIVE: To compare the difference in the efficacy on gait time cycle of ischemic stroke between yin-yang respiratory reinforcing and reducing needling technique (yin-yang needling) and the conventional acupuncture. METHODS: Sixty cases of ischemic stroke were randomized into a conventional acupuncture group and a yin-yang needling group, 30 cases in each one. The basic treatment (the control of blood pressure, blood sugar and blood lipid, the intravenous drops of ginkgo leaf extract and dipyridamole injection and vinpocetine injection) were applied in the two groups. Additionally, in the conventional acupuncture group, the acupoints of the Stomach Meridian of Foot-Yangming [Biguan (ST 31), Liangqiu (ST 34), Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), etc.] were selected and stimulated with the even needling technique. In the yin-yang needling group, the acupoints of yin meridians such as Zuwuli (LR 10), Xuehai (SP 10), Yinlingquan (SP 9) and Sanyinjiao (SP 6), etc. and the acupoints of yang meridians such as Biguan (ST 31), Liangqiu (ST 34) and Yanglingquan (GB 34), etc. were selected. The reducing manipulation of respiratory reinforcing and reducing technique was applied to the acupoints of yin meridians and the reinforcing manipulation was applied to the acupoints of yang meridians. The kinematics time parameters were determined and compared before and 4 weeks after treatment. RESULTS: After treatment, the differences in the gait cycle, the phase time of standing (%), the phase time of single support (%), the phase time of unilateral sway (%) on the affected (healthy) foot and phase time of double support (%) were significant as compared with those before treatment in the patients of the two groups (all P < 0.05), in which, the gait cycle (1.75 +/- 0.21 vs 2.02 +/- 0.37), the phase time of standing (%) on the affected (healthy) foot [(65.41 +/- 5.20)% vs (68.37 +/- 6.24)%, (70.99 +/- 6.47)% vs (74.51 +/- 5.19)%], the phase time of unilateral sway (%) on the affected (healthy) foot [(36.08 +/- 4.86)% vs (33.65 +/- 2.94)%, (31.04 +/- 3.41)% vs (26.77 +/- 2.67)%] and the phase time of double support (%) [(36.91 +/- 5.10)% vs (41.22 +/- 5.39)%] in the yin-yang needling group were improved much obviously after treatment as compared with those in the conventional acupuncture group. The differences in support phase time (%), single support phase time (%) and sway phase time (%) were significant between the affected limb and healthy limb of the two groups after treatment (all P < 0.05). CONCLUSION: Yin-yang respiratory reinforcing and reducing needling technique effectively improves hemiplegic gait movement cycle and walking function in patients of ischemic stroke, which is superior to the conventional acupuncture treatment.

[Impacts of yin-yang meridians acupuncture with respiratory reinforcing and reducing manipulation on lower limbs balance function in stroke patients].

OBJECTIVE: To observe the impact on lower limbs balance function in treatment of yin-yang meridians acupuncture with respiratory reinforcing and reducing manipulation involved in the patients of stroke by applying B-PHY balance function test training system so as to provide the objective evidence in treatment of stroke; with acupuncture. METHODS: One hundred patients were randomized into an observation group and a control group, 50 cases in each one. In the control group, the basic treatment was applied, without other relevant rehabilitation therapies associated. In the observation group, with the basic treatment as the control group's, the therapy of the yin-yang meridians acupuncutre with respiratory reinforcing and reducing manipulation was adopted. On the yin meridians, Zuwuli (LR 10), Xuehai (SP 10), Yinlingquan (SP 9), Sanyinjiao (SP 6) and the others were selected and stimulated with reducing manipulation achieved by the coordination of patient's respiration. On the yang meridians, Biguan (ST 31), Liangqiu (ST 34), Yanglingquan (GB 34) and the others were selected and stimulated with reinforcing manipulation achieved by the coordination of patient's respiration. The treatment was given once a day and for 28 days totally. Before treatment and in 28 days of treatment, B-PHY balance function test training system was used to determine the weight shift track parameters (track length, peripheral square, track length of per unit square, left-right offset and rectangle square), the weight shift track distance parameters [mean of X axle weight shift distance (Mean-X), mean of Y axle weight shift distance (Mean-Y), maximum of X axle weight shift distance (Max-X), maximum of Y axle weight shift distance (Max-Y), weight shift distance (LSKG), weight shift square (SSKG), square ratio of weight shift (LFS)], stability coefficient (SI) and weight distribution coefficient (WDI). RESULTS: After treatment, the differences in the weight shift track parameters, SI and WDI were significant as compared with those before treatment in the patients of the two groups (all P<0.01); while the differences in the weight shift distance parameters in the observation group were improved obviously after treatment as compared with those before treatment (all P<0.01), the differences of Mean-X, Max-Y and LFS in the control group were improved after treatment as compared with those before treatment (all P<0.01). Except SSKG, the improvements after treatment in the rest indices in the observation group were better than those in the control group (all P<0.05). CONCLUSION: The yin-yang meridians acupuncture with respiratry reinforcing and re- ducing manipulation effectively improves the lower limbs balance function in the patients of stroke.